Literature summary for 5.1.3.31 extracted from

Qi, Z.; Zhu, Z.; Wang, J.W.; Li, S.; Guo, Q.; Xu, P.; Lu, F.; Qin, H.M.
Biochemical analysis and the preliminary crystallographic characterization of D-tagatose 3-epimerase from Rhodobacter sphaeroides (2017), Microb. Cell Fact., 16, 193 .

Search for more literature for 5.1.3.31

Cloned(Commentary)

Cloned (Comment)	Organism
gene RSP_3671, sequence comparisons and phylogenetic tree, recombinant expression in Escherichia coli strain BL21(DE3)	Cereibacter sphaeroides

Protein Variants

Protein Variants	Comment	Organism
R118W	site-directed mutagenesis, the unique hydrogen bond between Arg118 and O4 of D-fructose is broken when Arg118 is mutated to Trp, the mutation improves the substrate recognition and activity of the enzyme. The mutant enzyme RsDTE_R118W shows decreased catalytic activity compared to the wild-type enzyme toward D-fructose, the kcat/Km for D-tagatose is about twofold higher than for D-psicose. Mutant R118W shows 1.5fold higher catalytic efficiency toward D-tagatose than the wild-type	Cereibacter sphaeroides

Inhibitors

Inhibitors	Comment	Organism	Structure
Ca2+	-	Cereibacter sphaeroides
Cu2+	high inhibition at 0.1 mM	Cereibacter sphaeroides
EDTA	-	Cereibacter sphaeroides
Fe2+	high inhibition at 0.1 mM	Cereibacter sphaeroides
Fe3+	-	Cereibacter sphaeroides
Ni2+	almost complete inhibition at 0.1 mM	Cereibacter sphaeroides
Zn2+	-	Cereibacter sphaeroides

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics	Cereibacter sphaeroides
0.98	-	D-tagatose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
78	-	D-fructose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
137.9	-	D-tagatose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
148.1	-	D-sorbose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
162.1	-	D-fructose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
192.9	-	D-psicose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
215.2	-	D-psicose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
359.4	-	D-sorbose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Co2+	dependent on, activates at 0.1 mM, metalloenzyme	Cereibacter sphaeroides
Mn2+	dependent on, activates at 0.1 mM, metalloenzyme, Mn2+ is the most effective activating divalent cation	Cereibacter sphaeroides
additional information	no effect on enzyme activity by Mg2+ at .1 mM	Cereibacter sphaeroides

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-fructose	Cereibacter sphaeroides	-	D-psicose	-	r
D-fructose	Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158	-	D-psicose	-	r
D-tagatose	Cereibacter sphaeroides	-	D-sorbose	-	r

Organism

Organism	UniProt	Comment	Textmining
Cereibacter sphaeroides	Q3IW04	-	-
Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158	Q3IW04	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-fructose	-	Cereibacter sphaeroides	D-psicose	-	r
D-fructose	best substrate for the recombinant wild-type enzyme	Cereibacter sphaeroides	D-psicose	-	r
D-fructose	-	Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158	D-psicose	-	r
D-fructose	best substrate for the recombinant wild-type enzyme	Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158	D-psicose	-	r
D-psicose	-	Cereibacter sphaeroides	D-fructose	-	r
D-psicose	-	Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158	D-fructose	-	r
D-sorbose	-	Cereibacter sphaeroides	D-tagatose	-	r
D-sorbose	-	Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158	D-tagatose	-	r
D-tagatose	-	Cereibacter sphaeroides	D-sorbose	-	r
D-tagatose	best substrate for the recombinant enzyme mutant R118W	Cereibacter sphaeroides	D-sorbose	-	r
additional information	RsDTE wild-type shows lower Michaelis-Menten constant (Km), lower turnover number (kcat), but higher catalytic efficiency (kcat/Km) values for D-fructose than for D-psicose. The kcat/Km for D-fructose is 5.5fold higher than for D-psicose, indicating that enzyme RsDTE highly catalyzes D-fructose, although it is a D-tagatose 3-epimerase	Cereibacter sphaeroides	?	-	?
additional information	RsDTE wild-type shows lower Michaelis-Menten constant (Km), lower turnover number (kcat), but higher catalytic efficiency (kcat/Km) values for D-fructose than for D-psicose. The kcat/Km for D-fructose is 5.5fold higher than for D-psicose, indicating that enzyme RsDTE highly catalyzes D-fructose, although it is a D-tagatose 3-epimerase	Cereibacter sphaeroides ATCC 17023 / 2.4.1 / NCIB 8253 / DSM 158	?	-	?

Synonyms

Synonyms	Comment	Organism
RsDTE	-	Cereibacter sphaeroides
RSP_3671	-	Cereibacter sphaeroides

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
35	-	-	Cereibacter sphaeroides

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
20	70	half-maximal activity at 20°C, near inactivation at 70°C	Cereibacter sphaeroides

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
8.3	-	D-sorbose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
15.36	-	D-sorbose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
20.47	-	D-psicose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
26.04	-	D-psicose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
41.05	-	D-fructose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
42.6	-	D-fructose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
48.31	-	D-tagatose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
51.25	-	D-tagatose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.5	-	-	Cereibacter sphaeroides

pH Range

pH Minimum	pH Maximum	Comment	Organism
5.5	11	activity range	Cereibacter sphaeroides

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the D-tagatose 3-epimerase (D-TE) family	Cereibacter sphaeroides
additional information	the O1 of substrate D-fructose forms hydrogen bonds with His192 and Glu162 to help the correct metal coordination of the substrate. The O2 forms hydrogen bonds with His192 and Arg221. Glu156 forms hydrogen bonds with O3, and Glu250 directs its OE2 atom to a hydrogen atom attached to C3. Because D-fructose has the same configurations of C1, C2 and C3 as D-tagatose, the interactions between D-fructose at the 1-, 2- and 3-positions and the enzyme are very similar to those in other DTE/DPE family enzymes. Residue R118 forms a hydrogen bond with O4 of D-fructose and may regulate the substrate specificity. The strengthened hydrophobic interaction may attribute to the recognition of D-tagatose, D-psicose, and D-sorbose. Enzyme homology modeling and structure comparisons, overview	Cereibacter sphaeroides
physiological function	the purified D-tagatose 3-epimerase from Rhodobacter sphaeroides catalyzes the epimerization of D-fructose to D-psicose at the C3 position	Cereibacter sphaeroides

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.02	-	D-sorbose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
0.1	-	D-sorbose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
0.1	-	D-psicose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
0.14	-	D-psicose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
0.25	-	D-fructose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
0.35	-	D-tagatose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides
0.52	-	D-tagatose	recombinant mutant R118W, pH 8.5, 35°C	Cereibacter sphaeroides
0.55	-	D-fructose	recombinant wild-type enzyme, pH 8.5, 35°C	Cereibacter sphaeroides

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Release 2023.1 (January 2023)